Bouilly K, Chaves R, Leitão A, Benabdelmouna A, Guedes-Pinto H
Institute for Biotechnology and Bioengineering, Centre of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, (IBB/CGB-UTAD), 5001-801 Vila Real, Portugal.
J Genet. 2008 Aug;87(2):119-25. doi: 10.1007/s12041-008-0018-2.
Chromosome identification is essential in oyster genomic research. Fluorescence in situ hybridization (FISH) offers new opportunities for the identification of oyster chromosomes. It has been used to locate satellite DNAs, telomeres or ribosomal DNA sequences. However, regarding chromosome identification, no study has been conducted with simple sequence repeats (SSRs). FISH was used to probe the physical organization of three particular SSRs, (GGAT)(4), (GT)(7) and (TA)(10) onto metaphase chromosomes of the Pacific oyster, Crassostrea gigas. Hybridization signals were observed in all the SSR probes, but the distribution and intensity of signals varied according to the oligonucleotide repeat. The intercalary, centromeric and telomeric bands were observed along the chromosomes, and for each particular repeat every chromosome pair presented a similar pattern, allowing karyotypic analysis with all the SSRs tested. Our study is the first in mollusks to show the application of SSR in situ hybridization for chromosome identification and karyotyping. This technique can be a useful tool for oyster comparative studies and to understand genome organization in different oyster taxa.
染色体识别在牡蛎基因组研究中至关重要。荧光原位杂交(FISH)为牡蛎染色体的识别提供了新的机遇。它已被用于定位卫星DNA、端粒或核糖体DNA序列。然而,关于染色体识别,尚未有针对简单序列重复(SSR)的研究。利用FISH技术将三个特定的SSR,即(GGAT)(4)、(GT)(7)和(TA)(10),定位到太平洋牡蛎(Crassostrea gigas)中期染色体上,以探究其物理组织情况。在所有SSR探针中均观察到杂交信号,但信号的分布和强度因寡核苷酸重复序列而异。沿着染色体观察到了居间带、着丝粒带和端粒带,并且对于每个特定的重复序列,每对染色体都呈现出相似的模式,从而使得对所有测试的SSR进行核型分析成为可能。我们的研究是软体动物中首次展示SSR原位杂交在染色体识别和核型分析中的应用。该技术可成为牡蛎比较研究以及理解不同牡蛎类群基因组组织的有用工具。